A real-time polymerase chain reaction assay for the detection of Mycoplasma agalactiae

AIM: To develop a real-time PCR for the detection of Mycoplasma agalactiae, using PCR primers targeting the ma-mp81 gene.

METHODS: A group of 15 M. agalactiae isolates, 21 other Mycoplasma spp. isolates and 21 other bacterial isolates was used in evaluation of the assay.

RESULTS: All M. agalactiae isolates were detected by the assay and none of the non-target isolates was amplified. The analytical detection limit of the assay was 10 fg of purified genomic DNA and 10⁴ cfu/ml milk inoculated with M. agalactiae. When applied to goat-milk samples collected from three herds free of M. agalactiae infection, the assay had a specificity of 100%.

CONCLUSIONS: The assay would be useful in a diagnostic laboratory, providing specific, sensitive and rapid detection of M. agalactiae.     

Efficiency of Repeated In Vivo Oocyte and Embryo Recovery After rhFSH Treatment in Rabbits

This study aims to assess the efficiency of in vivo oocyte and embryo recovery after a recombinant human FSH (rhFSH) treatment in rabbit does. Females were distributed in two experimental groups: donor does were treated with rhFSH (superovulation group) for 3 days prior to artificial insemination (embryo recovery) or ovulation induction (oocyte recovery) and does without treatment remained as the control group. Mature oocytes or embryos were collected with the laparoscopy technique 16 h after ovulation induction (oocytes) or 72 h after artificial insemination (embryos). Up to four recoveries were performed with each doe. Recovery efficiencies differed significantly between embryos (84%) and oocytes (58%). Yet, the recovery rates for the superovulation and control groups did not differ. The rhFSH group was associated with a significant increase (p < 0.05) in the number of oocytes and embryos recovered in comparison with the control group (10.2 ± 1.0 and 14.3 ± 1.2 vs 6.0 ± 2.7 and 8.4 ± 2.3 for oocytes and embryos, respectively). Results from this study indicate that repeated in vivo oocyte and embryo recovery from rhFSH superovulated does maximizes the number of oocytes or embryos collected from the same female.     

Efficiency of inspection procedures for the detection of tuberculous lesions in cattle

The sensitivity of the abattoir inspection procedure introduced for Australian export beef in 1976 was compared to a detailed necropsy procedure for the detection of tuberculous lesions in cattle. In a sample of cattle that were reactors to the tuberculin test, abattoir inspection failed to detect an estimated 47% of cattle with lesions. The detailed necropsy examination of cattle with lesions of tuberculosis identified 21 sites of infection compared with 13 to 18 in cattle examined by routine meat inspection procedures. Of the lesions detected during detailed necropsy, 15.9% did not involve the thoracic cavity or the medial retropharyngeal lymph nodes. The failure to detect lesions during abattoir inspection has its greatest significance in an animal with a single lesion. If the 245 cattle found with single lesions during detailed necropsy had been examined by abattoir inspection using the 1976 or the 1986 procedures, 0.8 and 8.9%, respectively, of these animals would not have been detected because the diseased tissues would not have been examined. If meat inspection is to provide an effective means of monitoring the level of bovine tuberculosis during the final stages of eradication, a procedure no less sensitive than that introduced in 1976 should be used.